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Mitochondrial fission determines cisplatin sensitivity in tongue squamous cell carcinoma through the BRCA1-miR-593-5p-MFF axis.

Fan S, Liu B, Sun L, Lv XB, Lin Z, Chen W, Chen W, Tang Q, Wang Y, Su Y, Jin S, Zhang D, Zhong J, Li Y, Wen B, Zhang Z, Yang P, Zhou B, Liang Q, Yu X, Zhu Y, Hu P, Chu J, Huang W, Feng Y, Peng H, Huang Q, Song E, Li J - Oncotarget (2015)

Bottom Line: The BRCA1-miR-593-5p-MFF axis also affected cisplatin sensitivity in vivo.Importantly, in a retrospective analysis of multiple centers, we further found that the BRCA1-miR-593-5p-MFF axis was significantly associated with cisplatin sensitivity and the survival of patients with TSCC.Together, our data reveal a model for mitochondrial fission regulation at the transcriptional and post-transcriptional levels; we also reveal a new pathway for BRCA1 in determining cisplatin sensitivity through the mitochondrial fission program.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.

ABSTRACT
Cisplatin has been widely employed as a cornerstone chemotherapy treatment for a wide spectrum of solid neoplasms; increasing tumor responsiveness to cisplatin has been a topic of interest for the past 30 years. Strong evidence has indicated that mitochondrial fission participates in the regulation of apoptosis in many diseases; however, whether mitochondrial fission regulates cisplatin sensitivity remains poorly understood. Here, we show that MFF mediated mitochondrial fission and apoptosis in tongue squamous cell carcinoma (TSCC) cells after cisplatin treatment and that miR-593-5p was downregulated in this process. miR-593-5p attenuated mitochondrial fission and cisplatin sensitivity by targeting the 3' untranslated region sequence of MFF and inhibiting its translation. In exploring the underlying mechanism of miR-593-5p downregulation, we observed that BRCA1 transactivated miR-593-5p expression and attenuated cisplatin sensitivity in vitro. The BRCA1-miR-593-5p-MFF axis also affected cisplatin sensitivity in vivo. Importantly, in a retrospective analysis of multiple centers, we further found that the BRCA1-miR-593-5p-MFF axis was significantly associated with cisplatin sensitivity and the survival of patients with TSCC. Together, our data reveal a model for mitochondrial fission regulation at the transcriptional and post-transcriptional levels; we also reveal a new pathway for BRCA1 in determining cisplatin sensitivity through the mitochondrial fission program.

No MeSH data available.


Related in: MedlinePlus

MFF regulates mitochondrial fission and apoptosis in TSCC cells after cisplatin treatmentA, Cisplatin induces mitochondrial fission with elevated MFF protein levels in Cal-27 and Scc-9 cells. Upper panel: MFF levels were analyzed via immunoblotting after cisplatin treatment. Lower panel: the quantification of cells with mitochondrial fission. #P < 0.05 versus no cisplatin treatment; *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment. B, Cytochrome c (CYTO c) distribution in mitochondria-enriched heavy membranes (HM) or the cytosol as detected via immunoblotting. C, D, E and F, Knockdown of MFF attenuated cisplatin-induced mitochondrial fission and apoptosis in Cal-27 and Scc-9 cells. Mitochondrial fission was detected via staining with MitoTracker Red. Scale bar equals 3 μm. Cell apoptosis was detected using TUNEL, flow cytometry, and caspase-3/7 activity assays. *P < 0.01 versus cisplatin alone; **P< 0.001 versus cisplatin alone. G, Cal-27 and Scc-9 cells transiently transfected with MFF expressing plasmids for 24 h were analyzed for MFF levels via immunoblotting. H, I, J and K, Mitochondrial fission and apoptosis were detected via staining with MitoTracker Red, flow cytometry, TUNEL, and caspase-3/7 activity assays. *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment.
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Figure 1: MFF regulates mitochondrial fission and apoptosis in TSCC cells after cisplatin treatmentA, Cisplatin induces mitochondrial fission with elevated MFF protein levels in Cal-27 and Scc-9 cells. Upper panel: MFF levels were analyzed via immunoblotting after cisplatin treatment. Lower panel: the quantification of cells with mitochondrial fission. #P < 0.05 versus no cisplatin treatment; *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment. B, Cytochrome c (CYTO c) distribution in mitochondria-enriched heavy membranes (HM) or the cytosol as detected via immunoblotting. C, D, E and F, Knockdown of MFF attenuated cisplatin-induced mitochondrial fission and apoptosis in Cal-27 and Scc-9 cells. Mitochondrial fission was detected via staining with MitoTracker Red. Scale bar equals 3 μm. Cell apoptosis was detected using TUNEL, flow cytometry, and caspase-3/7 activity assays. *P < 0.01 versus cisplatin alone; **P< 0.001 versus cisplatin alone. G, Cal-27 and Scc-9 cells transiently transfected with MFF expressing plasmids for 24 h were analyzed for MFF levels via immunoblotting. H, I, J and K, Mitochondrial fission and apoptosis were detected via staining with MitoTracker Red, flow cytometry, TUNEL, and caspase-3/7 activity assays. *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment.

Mentions: Cisplatin induced mitochondrial fission with elevated MFF protein levels (Figure 1A), but not elevated mRNA levels (Supplementary Figure S2A). Immunofluorescence microscopy revealed that MFF exhibited punctate localization in mitochondria and that mitochondria fragmentation occurred upon cisplatin treatment of TSCC cells (Supplementary Figure S2B). MFF knockdown attenuated the MFF protein upregulation (Supplementary Figure S2C) and partially inhibited the release of cytochrome c in the intermembrane space of mitochondria (Figure 1B) of cisplatin-treated cells. Cisplatin induced an alteration in the expression of FIS1, DRP1, MFN1, MFN2 and optic atrophy type I (OPA1); this alteration was not affected by MFF siRNA (Supplementary Figure S2C). Consequently, mitochondrial fission (Figure 1C) and the apoptosis of TSCC cells (Figure 1D-F) were attenuated by MFF siRNA. By contrast, enforced MFF expression led to mitochondrial fission and apoptosis (Figure 1G-K). These data suggest that MFF regulates mitochondrial fission and cisplatin sensitivity in TSCC cells.


Mitochondrial fission determines cisplatin sensitivity in tongue squamous cell carcinoma through the BRCA1-miR-593-5p-MFF axis.

Fan S, Liu B, Sun L, Lv XB, Lin Z, Chen W, Chen W, Tang Q, Wang Y, Su Y, Jin S, Zhang D, Zhong J, Li Y, Wen B, Zhang Z, Yang P, Zhou B, Liang Q, Yu X, Zhu Y, Hu P, Chu J, Huang W, Feng Y, Peng H, Huang Q, Song E, Li J - Oncotarget (2015)

MFF regulates mitochondrial fission and apoptosis in TSCC cells after cisplatin treatmentA, Cisplatin induces mitochondrial fission with elevated MFF protein levels in Cal-27 and Scc-9 cells. Upper panel: MFF levels were analyzed via immunoblotting after cisplatin treatment. Lower panel: the quantification of cells with mitochondrial fission. #P < 0.05 versus no cisplatin treatment; *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment. B, Cytochrome c (CYTO c) distribution in mitochondria-enriched heavy membranes (HM) or the cytosol as detected via immunoblotting. C, D, E and F, Knockdown of MFF attenuated cisplatin-induced mitochondrial fission and apoptosis in Cal-27 and Scc-9 cells. Mitochondrial fission was detected via staining with MitoTracker Red. Scale bar equals 3 μm. Cell apoptosis was detected using TUNEL, flow cytometry, and caspase-3/7 activity assays. *P < 0.01 versus cisplatin alone; **P< 0.001 versus cisplatin alone. G, Cal-27 and Scc-9 cells transiently transfected with MFF expressing plasmids for 24 h were analyzed for MFF levels via immunoblotting. H, I, J and K, Mitochondrial fission and apoptosis were detected via staining with MitoTracker Red, flow cytometry, TUNEL, and caspase-3/7 activity assays. *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment.
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Figure 1: MFF regulates mitochondrial fission and apoptosis in TSCC cells after cisplatin treatmentA, Cisplatin induces mitochondrial fission with elevated MFF protein levels in Cal-27 and Scc-9 cells. Upper panel: MFF levels were analyzed via immunoblotting after cisplatin treatment. Lower panel: the quantification of cells with mitochondrial fission. #P < 0.05 versus no cisplatin treatment; *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment. B, Cytochrome c (CYTO c) distribution in mitochondria-enriched heavy membranes (HM) or the cytosol as detected via immunoblotting. C, D, E and F, Knockdown of MFF attenuated cisplatin-induced mitochondrial fission and apoptosis in Cal-27 and Scc-9 cells. Mitochondrial fission was detected via staining with MitoTracker Red. Scale bar equals 3 μm. Cell apoptosis was detected using TUNEL, flow cytometry, and caspase-3/7 activity assays. *P < 0.01 versus cisplatin alone; **P< 0.001 versus cisplatin alone. G, Cal-27 and Scc-9 cells transiently transfected with MFF expressing plasmids for 24 h were analyzed for MFF levels via immunoblotting. H, I, J and K, Mitochondrial fission and apoptosis were detected via staining with MitoTracker Red, flow cytometry, TUNEL, and caspase-3/7 activity assays. *P < 0.01 versus no cisplatin treatment; **P < 0.001 versus no cisplatin treatment.
Mentions: Cisplatin induced mitochondrial fission with elevated MFF protein levels (Figure 1A), but not elevated mRNA levels (Supplementary Figure S2A). Immunofluorescence microscopy revealed that MFF exhibited punctate localization in mitochondria and that mitochondria fragmentation occurred upon cisplatin treatment of TSCC cells (Supplementary Figure S2B). MFF knockdown attenuated the MFF protein upregulation (Supplementary Figure S2C) and partially inhibited the release of cytochrome c in the intermembrane space of mitochondria (Figure 1B) of cisplatin-treated cells. Cisplatin induced an alteration in the expression of FIS1, DRP1, MFN1, MFN2 and optic atrophy type I (OPA1); this alteration was not affected by MFF siRNA (Supplementary Figure S2C). Consequently, mitochondrial fission (Figure 1C) and the apoptosis of TSCC cells (Figure 1D-F) were attenuated by MFF siRNA. By contrast, enforced MFF expression led to mitochondrial fission and apoptosis (Figure 1G-K). These data suggest that MFF regulates mitochondrial fission and cisplatin sensitivity in TSCC cells.

Bottom Line: The BRCA1-miR-593-5p-MFF axis also affected cisplatin sensitivity in vivo.Importantly, in a retrospective analysis of multiple centers, we further found that the BRCA1-miR-593-5p-MFF axis was significantly associated with cisplatin sensitivity and the survival of patients with TSCC.Together, our data reveal a model for mitochondrial fission regulation at the transcriptional and post-transcriptional levels; we also reveal a new pathway for BRCA1 in determining cisplatin sensitivity through the mitochondrial fission program.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.

ABSTRACT
Cisplatin has been widely employed as a cornerstone chemotherapy treatment for a wide spectrum of solid neoplasms; increasing tumor responsiveness to cisplatin has been a topic of interest for the past 30 years. Strong evidence has indicated that mitochondrial fission participates in the regulation of apoptosis in many diseases; however, whether mitochondrial fission regulates cisplatin sensitivity remains poorly understood. Here, we show that MFF mediated mitochondrial fission and apoptosis in tongue squamous cell carcinoma (TSCC) cells after cisplatin treatment and that miR-593-5p was downregulated in this process. miR-593-5p attenuated mitochondrial fission and cisplatin sensitivity by targeting the 3' untranslated region sequence of MFF and inhibiting its translation. In exploring the underlying mechanism of miR-593-5p downregulation, we observed that BRCA1 transactivated miR-593-5p expression and attenuated cisplatin sensitivity in vitro. The BRCA1-miR-593-5p-MFF axis also affected cisplatin sensitivity in vivo. Importantly, in a retrospective analysis of multiple centers, we further found that the BRCA1-miR-593-5p-MFF axis was significantly associated with cisplatin sensitivity and the survival of patients with TSCC. Together, our data reveal a model for mitochondrial fission regulation at the transcriptional and post-transcriptional levels; we also reveal a new pathway for BRCA1 in determining cisplatin sensitivity through the mitochondrial fission program.

No MeSH data available.


Related in: MedlinePlus